pAAV-hSyn-GRAB_CCK1.0

LOCUS       pAAV-hSyn-GRAB_CCK1.0           6611 bp DNA     circular SYN 23-JAN-2024
DEFINITION  Expresses the genetically-encoded fluorescent cholecystokinin (CCK) 
            sensor GRAB_CCK1.0 in neurons.
ACCESSION   .
VERSION     .
KEYWORDS    pAAV-hSyn-GRAB_CCK1.0
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6611)
  AUTHORS   Wang H, Qian T, Zhao Y, Zhuo Y, Wu C, Osakada T, Chen P, Chen Z, Ren
            H, Yan Y, Geng L, Fu S, Mei L, Li G, Wu L, Jiang Y, Qian W, Zhang L,
            Peng W, Xu M, Hu J, Jiang M, Chen L, Tang C, Zhu Y, Lin D, Zhou JN, 
            Li Y
  TITLE     A tool kit of highly selective and sensitive genetically encoded 
            neuropeptide sensors.
  JOURNAL   Science. 2023 Nov 17;382(6672):eabq8173. doi: 
            10.1126/science.abq8173. Epub 2023 Nov 17.
  PUBMED    37972184
REFERENCE   2  (bases 1 to 6611)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jul 9, 2026 from SnapGene 6.0.2
            https://www.biofeng.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Science."; 
            date: "2023-11-17"; volume: "382(6672):eabq8173. doi"; pages: " 
            10.1126/science.abq8173. Epub 2023 Nov 17"
FEATURES             Location/Qualifiers
     source          1..6611
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        131..574
                     /label=hSyn
     regulatory      581..590
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             587..2713
                     /codon_start=1
                     /label=CCK1.0
                     /translation="METDTLLLWVLLLWVPGSTGDTSLYKKVGTTGELLKLNRSVQGTG
                     PGPGASLCRPGAPLLNSSSVGNLSCEPPRIRGAGTRELELAIRITLYAVIFLMSVGGNM
                     LIIVVLGLSRRLRTVTNAFLLSLAVSDLLLAVACMPFTLLPNVMGTWIFGTVICKAVSY
                     LMGVSVSVSTLSLVAIALERYSAICRPGQARVWQTRSHAARVIVATWLLSGLLMVPYPV
                     YTVVQPVGPRVLQCVHRWPSARVRQTWSVLLLLLLFFIPGVVMAVAYGLISRELYLGLR
                     LPPGGTERRPNGLGPERSAGPGGAEAEPLPTQLNGAPGEPAPAGPRDTDALDLEEGGNV
                     YIKADKQKNGIKATFVIRHNIEDGGVQLASHYQQNTPIGDGPVLLPDNHYLSVQSKLSK
                     DPNEKRDHMVLLEFVTAAGITLGMDELYKGGTGGSMVRKGEELFTGVVPILVELDGDVN
                     GHKFSVHGEGEGDATLGKLTLKFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHD
                     FFKSAMPEGYIQERTIFFKDDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKL
                     EYNTGAGAKLLAKKRVVRMLLVIVVLFFLCWLPVYSANTWRAFDGPGAHRALSGAPISF
                     IHLLSYASACVNPLVYCFMHRRFRQACLETCARCCPRPPRARPRALPDEDPPTPSIASL
                     SRLSYTTISTLGPG"
     sig_peptide     587..649
                     /codon_start=1
                     /product="leader sequence from mouse immunoglobulin kappa 
                     light chain"
                     /label=Ig-kappa leader
                     /translation="METDTLLLWVLLLWVPGSTGD"
     protein_bind    650..674
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     1807..1828
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1934..1955)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2195..2214)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     misc_feature    2738..3326
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2791..2811)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3209..3220)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3358..3834
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3557..3576)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   3874..4014
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3874..4003
                     /label=AAV2 ITR
     rep_origin      4089..4544
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4176..4195)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4386..4407
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4561..4580)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4680..4702
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4740..4758)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4826..4930
                     /gene="bla"
                     /label=AmpR promoter
     CDS             4931..5791
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(5149..5168)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5962..6550
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6451..6470
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct gcggccgcac gcgtatgagg cggggtgggg gtgcctacct gacgaccgac
      181 cccgacccac tggacaagca cccaaccccc attccccaaa ttgcgcatcc cctatcagag
      241 agggggaggg gaaacaggat gcggcgaggc gcgtgcgcac tgccagcttc agcaccgcgg
      301 acagtgcctt cgcccccgcc tggcggcgcg cgccaccgcc gcctcagcac tgaaggcgcg
      361 ctgacgtcac tcgccggtcc cccgcaaact ccccttcccg gccaccttgg tcgcgtccgc
      421 gccgccgccg gcccagccgg accgcaccac gcgaggcgcg agataggggg gcacgggcgc
      481 gaccatctgc gctgcggcgc cggcgactca gcgctgcctc agtctgcggt gggcagcgga
      541 ggagtcgtgt cgtgcctgag agcgcagtcg agaaaccggt gccaccatgg agacagacac
      601 actcctgcta tgggtactgc tgctctgggt tccaggttcc actggtgaca caagtttgta
      661 caaaaaagtt ggcaccaccg gtgagctgct aaagctgaac cggagcgtgc agggaaccgg
      721 acccgggccg ggggcttccc tgtgccgccc gggggcgcct ctcctcaaca gcagcagtgt
      781 gggcaacctc agctgcgagc cccctcgcat tcgcggagcc gggacacgag aattggagct
      841 ggccattaga atcactcttt acgcagtgat cttcctgatg agcgttggag gaaatatgct
      901 catcatcgtg gtcctgggac tgagccgccg cctgaggact gtcaccaatg ccttcctcct
      961 ctcactggca gtcagcgacc tcctgctggc tgtggcttgc atgcccttca ccctcctgcc
     1021 caatgtcatg ggcacatgga tctttggcac cgtcatctgc aaggcggttt cctacctcat
     1081 gggggtgtct gtgagtgtgt ctacgctaag cctcgtggcc atcgcactgg agcggtacag
     1141 cgccatctgc cgaccaggcc aggcacgagt gtggcagacg cgctcccacg cggctcgcgt
     1201 gattgtagcc acgtggctgc tgtccggact actcatggtg ccctaccccg tgtacactgt
     1261 cgtgcaacca gtggggcctc gtgtgctgca gtgcgtgcat cgctggccca gtgcgcgggt
     1321 ccgccagacc tggtccgtac tgctgcttct gctcttgttc ttcatcccgg gtgtggttat
     1381 ggccgtggcc tacgggctta tctctcgcga gctctactta gggcttcgcc ttccgccggg
     1441 gggcaccgag cgcaggccca acggtctggg ccccgagcgc agcgcgggcc cggggggcgc
     1501 agaggccgaa ccgctgccca cccagctcaa cggcgcccct ggcgagcccg cgccggccgg
     1561 gccgcgcgac accgacgcgc tggacctgga ggagggagga aacgtctata tcaaggccga
     1621 caagcagaag aacggcatca aggcgacttt cgtgatccgc cacaacatcg aggacggcgg
     1681 cgtgcagctc gccagccact accagcagaa cacccccatc ggcgacggcc ccgtgctgct
     1741 gcccgacaac cactacctga gcgtgcagtc caaactttcg aaagacccca acgagaagcg
     1801 cgatcacatg gtcctgctgg agttcgtgac cgccgccggg atcactctcg gcatggacga
     1861 gctgtacaag ggcggtaccg gagggagcat ggtgagaaag ggcgaggagc tgttcaccgg
     1921 ggtggtgccc atcctggtcg agctggacgg cgacgtaaac ggccacaagt tcagcgtgca
     1981 cggcgagggt gagggcgatg ccaccttggg caagctgacc ctgaagttca tctgcaccac
     2041 cggcaagctg cccgtgccct ggcccaccct cgtgaccacc ctgacctacg gcgtgcagtg
     2101 cttcagccgc taccccgacc acatgaagca gcacgacttc ttcaagtccg ccatgcccga
     2161 aggctacatc caggagcgca ccatcttctt caaggacgac ggcaactaca agacccgcgc
     2221 cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc gagctgaagg gcatcgactt
     2281 caaggaggac ggcaacatcc tggggcacaa gctggagtac aacaccggag caggcgcaaa
     2341 gctgctggct aagaagcgcg tggtgcgaat gttgctggtg atcgttgtgc ttttttttct
     2401 gtgttggttg ccagtttata gtgccaacac gtggcgcgcc tttgatggcc cgggtgcaca
     2461 ccgagcactc tcgggtgctc ctatctcctt cattcacttg ctgagctacg cctcggcctg
     2521 tgtcaacccc ctggtctact gcttcatgca ccgtcgcttt cgccaggcct gcctggaaac
     2581 ttgcgctcgc tgctgccccc ggcctccacg agctcgcccc agggctcttc ccgatgagga
     2641 ccctcccact ccctccattg cttcgctgtc caggcttagc tacaccacca tcagcacact
     2701 gggccctggc tgagaattcg atatcaagct tatcgataat caacctctgg attacaaaat
     2761 ttgtgaaaga ttgactggta ttcttaacta tgttgctcct tttacgctat gtggatacgc
     2821 tgctttaatg cctttgtatc atgctattgc ttcccgtatg gctttcattt tctcctcctt
     2881 gtataaatcc tggttgctgt ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg
     2941 cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt tggggcattg ccaccacctg
     3001 tcagctcctt tccgggactt tcgctttccc cctccctatt gccacggcgg aactcatcgc
     3061 cgcctgcctt gcccgctgct ggacaggggc tcggctgttg ggcactgaca attccgtggt
     3121 gttgtcgggg aaatcatcgt cctttccttg gctgctcgcc tatgttgcca cctggattct
     3181 gcgcgggacg tccttctgct acgtcccttc ggccctcaat ccagcggacc ttccttcccg
     3241 cggcctgctg ccggctctgc ggcctcttcc gcgtcttcgc cttcgccctc agacgagtcg
     3301 gatctccctt tgggccgcct ccccgcatcg ataccgagcg ctgctcgaga gatctacggg
     3361 tggcatccct gtgacccctc cccagtgcct ctcctggccc tggaagttgc cactccagtg
     3421 cccaccagcc ttgtcctaat aaaattaagt tgcatcattt tgtctgacta ggtgtccttc
     3481 tataatatta tggggtggag gggggtggta tggagcaagg ggcaagttgg gaagacaacc
     3541 tgtagggcct gcggggtcta ttgggaacca agctggagtg cagtggcaca atcttggctc
     3601 actgcaatct ccgcctcctg ggttcaagcg attctcctgc ctcagcctcc cgagttgttg
     3661 ggattccagg catgcatgac caggctcagc taatttttgt ttttttggta gagacggggt
     3721 ttcaccatat tggccaggct ggtctccaac tcctaatctc aggtgatcta cccaccttgg
     3781 cctcccaaat tgctgggatt acaggcgtga accactgctc ccttccctgt ccttctgatt
     3841 ttgtaggtaa ccacgtgcgg accgagcggc cgcaggaacc cctagtgatg gagttggcca
     3901 ctccctctct gcgcgctcgc tcgctcactg aggccgggcg accaaaggtc gcccgacgcc
     3961 cgggctttgc ccgggcggcc tcagtgagcg agcgagcgcg cagctgcctg caggggcgcc
     4021 tgatgcggta ttttctcctt acgcatctgt gcggtatttc acaccgcata cgtcaaagca
     4081 accatagtac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag
     4141 cgtgaccgct acacttgcca gcgccttagc gcccgctcct ttcgctttct tcccttcctt
     4201 tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt
     4261 ccgatttagt gctttacggc acctcgaccc caaaaaactt gatttgggtg atggttcacg
     4321 tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt
     4381 taatagtgga ctcttgttcc aaactggaac aacactcaac tctatctcgg gctattcttt
     4441 tgatttataa gggattttgc cgatttcggt ctattggtta aaaaatgagc tgatttaaca
     4501 aaaatttaac gcgaatttta acaaaatatt aacgtttaca attttatggt gcactctcag
     4561 tacaatctgc tctgatgccg catagttaag ccagccccga cacccgccaa cacccgctga
     4621 cgcgccctga cgggcttgtc tgctcccggc atccgcttac agacaagctg tgaccgtctc
     4681 cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga gacgaaaggg
     4741 cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt cttagacgtc
     4801 aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca
     4861 ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa
     4921 aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt
     4981 ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca
     5041 gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag
     5101 ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc
     5161 ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca
     5221 gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt
     5281 aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct
     5341 gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt
     5401 aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga
     5461 caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact
     5521 tactctagct tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc
     5581 acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga
     5641 gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt
     5701 agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga
     5761 gataggtgcc tcactgatta agcattggta actgtcagac caagtttact catatatact
     5821 ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga
     5881 taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt
     5941 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca
     6001 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct
     6061 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta
     6121 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
     6181 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
     6241 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
     6301 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
     6361 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
     6421 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
     6481 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
     6541 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt
     6601 tgctcacatg t
//